JP2002090368A - Device and method for manufacturing micro-array - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a device and a method for manufacturing a micro-array with a uniform spot diameter even if foam is entrained in a flow passage for discharging a DNA solution. SOLUTION: In this micro-array manufacturing device 1 manufacturing a micro-array by discharging a very small amount of DNA solution containing a probe, which can be specifically bonded to a target substance, onto a membrane filter 74, a relative positional relationship between a flow passage member 31 holding the DNA solution inside its flow passage 34 and having a nozzle 35 in one end of the flow passage 34 and the membrane filter 74 is varied by means of an XYZ robot 72, and when the flow passage member 31 is moved forward/backward along the discharge direction (Z-direction) by means of a layered piezoelectric element 29, the DNA solution held in the flow passage 34 of the flow passage member 31 is discharged in a matrix form from the nozzle 35 onto the membrane filter 74 by means of inertial force.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a microarray manufacturing apparatus for manufacturing a microarray in which liquid spots including probes are arranged in a matrix at high density on a substrate, and a microarray using the microarray manufacturing apparatus. The present invention relates to a method for manufacturing a microarray to be manufactured.

[0002]

2. Description of the Related Art As a prior art of a method for manufacturing a microarray, for example, there is one disclosed in Japanese Patent Application Laid-Open No. 11-187900. In this conventional technique, a spot containing a probe is formed on a solid phase by attaching a liquid sample containing a probe as droplets on a solid phase (solid substrate) by an inkjet method. Hereinafter, FIG.
An outline of a method for manufacturing a microarray according to the above-described conventional technique will be described with reference to FIG.

In FIG. 10, reference numeral 101 denotes a liquid supply system (nozzle) which holds a liquid containing a probe (eg, a nucleic acid probe) as a discharge liquid so as to be able to discharge the liquid, and 103 denotes a solid phase (to which the nucleic acid probe is to be bound). For example, a transparent glass plate) 105 is a type of ink jet head, and is a bubble jet (registered trademark) head having a mechanism for applying thermal energy to the liquid and discharging the liquid.
Reference numeral 04 denotes a liquid containing a nucleic acid probe discharged from the bubble jet head 105.

In the bubble jet head 105 shown in FIG. 11, reference numeral 107 denotes a liquid containing a nucleic acid probe to be discharged, and reference numeral 117 denotes a substrate portion having a heat generating portion for applying discharge energy to the liquid. This substrate portion 117 is provided with a protective film 10 made of silicon oxide or the like.
9 and an electrode 111-1 formed of aluminum or the like.
111-2, a heating resistor layer 113 formed of a nichrome wire or the like, a heat storage layer 115, and a support 116 formed of alumina or the like having good heat dissipation.

The liquid 107 containing the nucleic acid probe reaches a discharge orifice (discharge port) 119 and forms a meniscus 121 under a predetermined pressure. When an electric signal is applied to the electrodes 111-1 and 111-2 in this state, a region (foaming region) indicated by reference numeral 123 generates heat rapidly, and bubbles are generated in the liquid 107 in contact with this region. The meniscus is discharged by the pressure of the bubble, and the liquid 107 is discharged from the orifice 119. The discharged liquid 107 flies toward the surface of the solid phase 103.

[0006] The amount of liquid that can be ejected using the bubble jet head having the above-described structure depends on the size of the nozzle and the like, but can be controlled to, for example, about 4 to 50 picoliters. This is extremely effective as a means for arranging nucleic acid probes at high density. Note that the above-mentioned prior art publication states that a piezo jet head that discharges a liquid in a nozzle by using the vibration pressure of a piezo element can be used instead of the above-described bubble jet head. .

[0007]

The above prior art has the advantage that a minute amount of probe sample can be arranged on a solid substrate with high accuracy and high density. Bubbles may be mixed in the ejection head
If bubbles are mixed in the ejection head, the amount of ejected liquid may change or the ejection of the liquid may not be possible. In this case, there is a problem that the spot diameter of the microarray becomes non-uniform. That is, in the above-described conventional technique, bubbles may be mixed into the flow path due to the exchange of the liquid sample or the vaporization of dissolved oxygen in the liquid sample, and when the bubbles are mixed, the pressure for discharging the liquid may be reduced. Is attenuated by the air bubbles mixed therein, and the amount of the liquid to be ejected changes.

An object of the present invention is to provide a microarray manufacturing apparatus and a manufacturing method capable of manufacturing a microarray having a uniform spot diameter even when bubbles are mixed in a flow path for discharging a liquid containing a probe. And

[0009]

According to a first aspect of the present invention, a small amount of a liquid containing a probe capable of specifically binding to a target substance is discharged onto a substrate. A microarray manufacturing apparatus that manufactures a microarray by holding the liquid in a flow path inside the flow path member and having a discharge port at one end of the flow path; and moving the flow path member back and forth along a discharge direction. And a relative moving means for changing a relative positional relationship between the substrate and the flow path member in a plane orthogonal to the discharge direction, and moving the flow path member in the discharge direction by the drive means. In this way, the liquid held in the flow path member is discharged from the discharge port.

To achieve the above object, a second invention according to claim 2 is a liquid container for supplying the liquid to the flow path member, wherein the liquid surface of the liquid is opened to the atmosphere. Further comprising a liquid container that accommodates the liquid container, the other end of the flow path of the flow path member is an open end, the open end is configured to be immersed in the liquid in the liquid container, and from the liquid container by capillary action. The liquid is supplied into the flow path member.

According to a third aspect of the present invention, the relative moving means is configured to change a relative positional relationship between the substrate and the flow path member by moving the substrate. And

According to a fourth aspect of the present invention, a microarray is manufactured by using the microarray manufacturing apparatus according to any one of the first to third aspects.

In the first invention, a liquid containing a probe capable of specifically binding to a target substance is held in a flow path therein, and a flow path member having a discharge port at one end of the flow path is driven by a driving means. By moving forward and backward along the discharge direction, when the flow path member moving in the direction to advance along the discharge direction stops, inertial force acts on the liquid in the flow path member, and A flow is generated in the liquid in the discharge direction, and a minute amount of droplets is discharged from the discharge port onto the substrate by the flow. The discharge of such a minute amount of liquid droplets is repeated by changing the relative positional relationship between the substrate and the flow path member in a plane perpendicular to the discharge direction by the relative moving means, so that the liquid is moved to a desired position on the substrate. Is discharged, and a microarray is manufactured.

According to the first aspect of the present invention, the inertial force acting on the liquid does not change its size even when bubbles are present in the flow path. It is possible to discharge at a constant speed. for that reason,
The accuracy of the discharge amount and the discharge position of the liquid discharged on the substrate can be improved. Further, since the flow path member holds the liquid in the flow path inside the flow path member, the pressure applied to the liquid by the forward / backward movement of the flow path member increases near the discharge port, so that the The discharge speed can be increased.

In the second aspect, the other end of the flow path provided inside the flow path member and having a discharge port at one end is an open end, and this open end is specifically bound to a target substance. A liquid container for supplying a liquid containing a possible probe to the flow channel member, wherein the liquid container is configured to be immersed in a liquid in a liquid container that accommodates the liquid so that the liquid surface is open to the atmosphere. Due to the phenomenon, the liquid is supplied from the liquid container into the flow path member, and the liquid is held in the flow path inside the liquid flow path member. By moving the flow path member forward and backward along the discharge direction by the driving means, when the flow path member moving in the direction proceeding along the discharge direction stops, the inertia force is applied to the liquid in the flow path member. As a result, a flow is generated in the liquid in the flow path in the discharge direction, and a minute amount of droplets is discharged from the discharge port onto the substrate by the flow. The discharge of such a minute amount of liquid droplets is repeated by changing the relative positional relationship between the substrate and the flow path member in a plane perpendicular to the discharge direction by the relative moving means, so that the liquid is moved to a desired position on the substrate. Is discharged, and a microarray is manufactured.

According to the second aspect of the present invention, the inertial force acting on the liquid does not change its size even when bubbles are present in the flow path. It is possible to discharge at a constant speed. for that reason,
The accuracy of the discharge amount and the discharge position of the liquid discharged on the substrate can be improved. Further, since the flow path member holds the liquid in the flow path inside the flow path member, the pressure applied to the liquid by the forward / backward movement of the flow path member increases near the discharge port, so that the The discharge speed can be increased. Furthermore, since a liquid supply mechanism for introducing the liquid into the flow path in the flow path member is not required, the size of the apparatus can be reduced and the apparatus cost can be reduced.

According to the third aspect, when changing the relative positional relationship between the substrate and the flow path member by the relative moving means, the substrate is moved, but the flow path member is not moved. When the positional relationship is changed, there is no possibility that the meniscus position of the discharge port of the flow path member fluctuates and affects the discharge position accuracy of the liquid.

According to the fourth aspect, the microarray is manufactured by using the microarray manufacturing apparatus according to any one of the first to third aspects. Therefore, the manufactured microarray is included in the first to third aspects described above. Has the corresponding effect.

[0019]

Embodiments of the present invention will be described below in detail with reference to the drawings. FIG. 1 is a front view showing the configuration of the microarray manufacturing apparatus according to the first embodiment of the present invention.
FIGS. 2A and 2B are a top view and a detailed view of a part B of the microarray manufacturing apparatus according to the first embodiment of the present invention, respectively. FIG. 3 shows the microarray manufacturing apparatus according to the first embodiment. FIGS. 4A to 4E are perspective views of the mounted liquid discharge unit, and FIGS. 4A to 4E are views for explaining a liquid discharge operation by the liquid discharge head of the liquid discharge unit of the first embodiment, and FIGS. FIG. 3 is a diagram illustrating a drive waveform of a laminated piezoelectric element of the liquid ejection head according to the first embodiment.

First, the schematic configuration of the microarray manufacturing apparatus 1 of the present embodiment is shown in FIGS. 1, 2 (a) and 2 (b) and FIG.
This will be described below. In the microarray manufacturing apparatus 1 of the present embodiment, an XYZ robot capable of moving a liquid discharge unit 71 including eight liquid discharge heads 20 shown in FIG. 3 in X, Y, and Z directions (left, right, front, back, and up and down). (Relative moving means) 72, and each liquid ejection head 20 is connected to a syringe piston pump via a Teflon (registered trademark) pipe 24 (only one at the right end is shown, and the others are not shown). 25.

The syringe piston pump 25 is connected to another pipe to the liquid supply tank 26 in addition to the Teflon pipe 24. With this other pipe, the syringe piston pump 25 connects the solenoid valve 27 and the water supply pump 28. Via a liquid supply tank 26. The liquid ejection head 20, the syringe piston pump 25, and the liquid supply tank 26 are installed at substantially the same height. The liquid supply tank 26 contains cleaning water or degassed ion-exchanged water, and the water supply pump 28 fills and supplies cleaning water to each pipe 24, the syringe piston pump 25, and the liquid discharge head 20. ing.

Above the liquid discharge head 20, a laminated piezoelectric element 29 as a driving means is provided. One end (the upper end in the figure) of the laminated piezoelectric element 29 in the displacement direction is fixed to the gantry 30, and the other end (the lower end in the figure) is fixed to the flow path member 31. The multilayer piezoelectric element 29 moves the flow path member 31 forward and backward in the discharge direction (Z direction) by the displacement. The flow path member 31 includes a liquid inlet 33, a flow path 34, and a nozzle 35, and the flow path 34 includes a straight portion 36 and a tapered portion 37, as shown in a partial cross-sectional view at the right end of FIG. The liquid inlet 33 is connected to the syringe piston pump 25 via the Teflon pipe 24. The approximate size of the flow path 34 is such that the straight portion 36 has a diameter of φ0.5 mm to φ4 mm.
And the length is 2 mm to 15 mm. The tapered portion 37 is formed from the straight portion 36 toward the nozzle 35, and has a taper angle of 10 degrees to 45 degrees. The diameter of the nozzle 35 is φ0.03 mm to φ0.15 mm, and the length is 0.05 mm to 1 mm.

A water-repellent layer made of a fluorine-based material that is a low surface energy substance is provided on the end face and the outer peripheral face of the nozzle 35. Since the space between the laminated piezoelectric element 29 and the flow path 34 is formed as a rigid body, the displacement of the laminated piezoelectric element 29 does not change the volume of the flow path 34. Also, since one end of the laminated piezoelectric element 29 is fixed to the gantry 30,
The entire flow path member 31 moves according to the displacement of the laminated piezoelectric element 29. A voltage having a desired waveform as described later is applied to the laminated piezoelectric element 29 from a drive circuit (not shown) by a lead wire or a flexible substrate.

As shown in FIG. 2 (a), the sample container 73 has a total of 96 holes of 8 rows × 12 columns, and six samples are placed on the base. Each sample container 73 contains a DNA solution containing a fluorescently labeled single-stranded DNA probe in the hole. Liquid ejection unit 7
One liquid ejection head 20 is arranged at the same pitch as the pitch of the eight rows of holes of the sample container 73. Numeral 74 denotes a porous membrane filter used as a substrate, on which fine droplets of a DNA solution are deposited by a method described later.
They are arranged in a dimensional matrix. Membrane filter 74
Has a size of 5 mm × 5 mm to 20 mm × 20 mm (for example, about 8 mm × 8 mm), and a plurality of sheets are suction-fixed on the movable stage 75 by a suction device (not shown).
In the present embodiment, as shown in the detailed view of FIG.
It is assumed that 28 membrane filters 74 are installed.

A cleaning tank 76 shown in FIG. 2 cleans the liquid discharge unit 71 using the cleaning tank 76. The liquid ejection unit 71 is moved between the positions of the cleaning tank 76, the sample container 73, and the membrane filter 74 by the XYZ robot 72, and is stopped at each position.

Next, the operations during the suction operation and the discharge operation when the microarray is manufactured by the microarray manufacturing apparatus 1 of the present embodiment will be described with reference to FIGS.
This will be described with reference to FIG. First, the suction operation will be described.
The liquid discharge unit 71 is moved to the upper part of the cleaning tank 76, and the nozzle 35 of the flow path member 31 of each liquid discharge head 20 is immersed in the cleaning tank 76 by 1 mm to 2 mm, and then the electromagnetic valve 27 is opened. The wash water in the liquid supply tank 26 is sent to the flow channel by the water feed pump 28, and
And the outer peripheral surface and the end surface of the nozzle 35 are washed with washing water. While the washing water is being supplied, the syringe piston pump 25 is moved to the middle point,
As a result, the syringe piston pump 25 is filled with cleaning water. In this state, after closing the electromagnetic valve 27, the liquid discharge head 20 is raised above the cleaning tank 76. Thereafter, the piston of the syringe piston pump 25 is moved upward by a predetermined amount, and the washing water is discharged from the nozzle 35. Thereafter, the piston of the syringe piston pump 25 is moved to the middle point again, and a predetermined amount of air is sucked into the flow path 34.

Next, the liquid discharge unit 71 is connected to the sample container 7.
3 to move the respective liquid ejection heads 20
The DNA solution in the hole in the sample container 73 is sucked into the flow channel 34 from the nozzle 35 of the flow channel member 31. The DNA solution sucked into the flow path is applied to the air layer 3 shown in FIG.
8 is separated from the washing water. After the suction of the DNA solution is completed, the liquid discharge unit 71 is moved onto the membrane filter 74 to stop positioning, and the relative positional relationship between the membrane filter 74 and the liquid discharge unit 71 in the XY plane is changed. While finely adjusting the ejection position, the DNA solution is ejected onto the membrane filter 74 by a method described later.

Next, the discharge operation will be described. While a voltage having a waveform as shown in FIG. 5 is applied to the laminated piezoelectric element 29 of the liquid discharge head 20, t <t1 in FIG.
Assuming that the vertical position of the tip of the nozzle 35 of the flow path member 31 when = E0 is A, t1 <t <t2 in FIG.
Liquid discharge head 2
0 is slowly displaced downward in the figure, and t in FIG.
Immediately before = t2, a displacement corresponding to the voltage E1 occurs in the laminated piezoelectric element 29, and the displacement causes the tip of the nozzle 35 to descend to the position B. The position A and the position B
Is, for example, 1 to 10 μm.

Thereafter, at t = t2 in FIG. 4D, the voltage E instantaneously decreases to E0. With the rapid decrease of the voltage E, the flow path member 31 of the liquid discharge head 20 changes from descending to ascending and rapidly moves upward in the drawing. This flow path member 3
When the descending flow path member 31 stops at the position B during the reciprocating movement along the discharge direction (Z direction) 1, the DNA solution in the flow path 36 of the flow path member 31 is moved downward in the drawing. The inertial force acts to generate a downward flow. Due to this flow, the pressure at the distal end of the flow path 36 increases, and a droplet of the DNA solution is discharged onto the membrane filter 74 from the end face of the nozzle 35. In this case, the amount of the liquid to be discharged varies depending on the nozzle diameter, the physical property value of the DNA solution, the drive voltage waveform, and the like, but is about 0.1 nl to 0.3 μL. Thereafter, at t> t2 in FIG. 4E, the liquid ejection head 20 returns to the initial position.

The diameter of the DNA solution droplet discharged as described above is about 50 μm to 200 μm, and such a droplet is placed on the membrane filter 74 by about 150 μm.
Discharge at intervals of up to 400 μm. In this case, the eight liquid ejection heads 20 separate the respective membrane filters 74 from each other.
After simultaneously discharging the DNA solution on the upper side, the liquid discharge unit 71 is moved again to the upper part of the washing tank 76, where the flow path 34
The DNA solution remaining inside is discharged, and the channel 34 and the outer peripheral surface are washed. Thereafter, the XYZ robot 72
By changing the relative positional relationship between the membrane filter 74 and the flow path member 31 of the liquid ejection head 20 in the Y plane, the above-described suction operation, ejection operation, and washing operation are repeated, and the fine droplets of the DNA solution are placed on the membrane filter 74. Are discharged in a two-dimensional matrix at intervals of 75 μm to 400 μm. At this time, since the membrane filter 74 is porous, the discharged DNA solution permeates the membrane filter 74 and is retained. Thus, a DNA probe array which is a microarray is prepared.

According to the present embodiment, the liquid discharge head 20
The DNA solution is ejected from the nozzle 35 at the tip of the flow path member 31 by the inertial force, and the inertia force does not change even if bubbles exist in the flow path 34.
A constant amount of the DNA solution is always discharged at a constant speed irrespective of the presence or absence of bubbles in the sample 4, and a DNA probe array having a uniform spot diameter can be manufactured with high discharge position accuracy. Further, since the pressure at the distal end of the flow path 36 increases due to the forward and backward movement of the flow path member 31 of the liquid discharge head 20, the discharge speed of the DNA solution can be increased.

In this embodiment, a single-stranded DNA probe is used as a sample for the microarray. However, the present invention is not limited to this. Examples of the sample constituting the array include a ligand, a receptor for the ligand,
Oligo or polypeptide having a specific amino acid sequence, various enzymes, various proteins, antigens and antibodies, DNA or RNA having a specific base sequence, PNA or the like may be used.

In this embodiment, the porous membrane filter 74 is used as the substrate. However, the present invention is not limited to this. For example, a slide glass, a silicon substrate, a plastic plate, a ceramic plate, or the like may be used. Good. However, in order to prevent contamination between adjacent probes, it is desirable that the probe discharged onto the substrate be fixed at a predetermined position. A functional group capable of reacting with each other may be bonded to both the substrate and the substrate. Some of these methods have already been published,
For example, a thiol (-SH) group is bonded to the end of the probe, and the substrate surface is treated so as to have a maleimide group, whereby the thiol group of the probe supplied to the substrate surface and the maleimide group on the substrate surface are treated. May be reacted to fix the probe on the substrate. Alternatively, an epoxy group may be introduced on the substrate, and the epoxy group may be reacted with the amino group at the terminal of the probe.

FIGS. 6 and 7 show the second embodiment of the present invention, respectively.
3A and 3B are a front view and a top view showing the configuration of the microarray manufacturing apparatus according to the embodiment, and the same parts as those in the first embodiment are denoted by the same reference numerals. In the microarray manufacturing apparatus 2 of the present embodiment, the membrane filter 74 is XY
A plurality of sheets are suction-fixed to the lower surface of a filter base 77 supported by the Z robot 72, and the vertical relationship of the membrane filter 74 is reversed from that of the first embodiment. FIG.
Reference numeral 78 denotes a discharge unit in which a plurality of liquid discharge heads 79 and a sample container 73 are integrated. Hereinafter, the discharge unit 7
8 will be described in detail with reference to FIG.

In FIG. 8, a sample container 73 is a liquid container for supplying a DNA solution to the flow path member 82,
Solution A is stored such that the liquid surface is open to the atmosphere. This sample container 73 has a total of 96 holes of 8 rows × 12 columns, and each hole has a single-stranded fluorescently labeled D
A DNA solution containing an NA probe is contained therein, and is fixed at a predetermined position on the unit base 80. Reference numeral 81 denotes a discharge head base serving as a base of the liquid discharge head 79, which has a U-shaped cross-sectional shape, and is fixedly fastened to the unit base 80 with screws so as to cover the sample container 73.

As shown in the sectional view of FIG. 9A, the liquid discharge head 79 includes a flow path member 82, a laminated piezoelectric element 29 which is a driving means for driving the flow path member 82, and a laminated piezoelectric element 29. Member 8 for rigidly connecting the flow path member 82 and the flow path member 82
3. A flow path 84 is formed inside the flow path member 82, and a nozzle 85 is formed at an upper end of the flow path 84 of the flow path member 82. A straight portion 87 is formed from the nozzle 85 via a tapered portion 86.
The inner surface of the flow channel 84 is coated with titanium oxide to provide a hydrophilic surface. Channel member 82
The lower end 82a of the flow path 84 is an open end, and since the lower end 82a is immersed in the DNA solution inside the hole of the sample container 73, the DNA solution in each hole of the sample container 73 is a capillary. It is sucked and held in the flow path 84 by the phenomenon. Since the surface energy of the flow channel 84 is particularly high due to titanium oxide, the DNA solution is sucked up by the flow channel 84 by capillary action, and fills the entire flow channel 84.

One end (lower end in the drawing) of the laminated piezoelectric element 29 in the polarization direction is adhered and fixed on the discharge head base 81, and the other end (the upper end in the drawing) is fixed to the flow path member 82 via the connecting member 83. In addition, when the laminated piezoelectric element 29 expands and contracts, the entire flow path member 82 moves forward and backward in the discharge direction (vertical direction in the drawing; Z direction). One ejection unit 78 has a total of 96 liquid ejection heads 79.
Are provided, and four such discharge units 78 are arranged at predetermined positions of the base 88. A desired voltage is applied to each laminated piezoelectric element 29 from a drive circuit (not shown) via a connector and a lead wire.

The membrane filter 74 is moved and arranged on each discharge unit 78 by the XYZ robot 72, whereby the relative positional relationship between the membrane filter 74 and the liquid discharge head 79 of each discharge unit 78 in the XY plane is adjusted. The distance between the membrane filter 74 and the nozzle 85 of the liquid ejection head 79 is approximately 0.5
mm to 2 mm.

Next, the operations during the suction operation and the discharge operation when the microarray is manufactured by the microarray manufacturing apparatus 2 of the present embodiment will be described with reference to FIGS.
First, the suction operation will be described. In FIG. 6, first, the XYZ robot 72 controls the membrane filter 7.
4 is moved to a position about 1 mm above a desired liquid ejection head 79 and positioned and fixed. Next, the drive voltage waveform of FIG. 5 described above is applied to the laminated piezoelectric element 29 of the liquid ejection head 79, and the DNA held in the flow path 84 from the nozzle 85 by the same operation as in the first embodiment. The solution is discharged as fine droplets toward the membrane filter 74 disposed above.

At this time, the flying speed of the discharged droplet is reduced by the influence of gravity, but the speed of the discharged droplet is 3 m / s with respect to the distance (for example, about 1 mm) between the nozzle 85 and the membrane filter 74. Since the speed is sufficiently high at 10 to 10 m / s, the ejected droplets surely land on the membrane filter 74. The size of the droplet of the DNA solution to be discharged is from φ30 μm to φ300 μm in diameter. Since the membrane filter 74 is porous, the discharged DNA solution permeates and is retained in the membrane filter 74. Thereafter, while moving the membrane filter 74 in a two-dimensional matrix in the XY directions shown in FIG. 7, the DNA solution is discharged from the liquid discharge head 79 at each position,
By arranging minute spots of the DNA solution in a matrix and at a high density on the membrane filter 74,
Create a probe array.

If the procedure for moving the membrane filter 74 and the discharge timing of the liquid discharge head 79 are determined in advance, the above operation can be performed automatically. The ejection unit 78 can be repeatedly used by disassembling and cleaning the sample container 73 and the ejection head base 81. If a plurality of ejection units 78 are prepared, cleaning of the ejection units and creation of the probe array can be performed simultaneously.

According to the present embodiment, the following effects are obtained in addition to the effects of the first embodiment. That is, in the present embodiment, the end opposite to the end provided with the nozzle 85 of the flow path 84 in the flow path member 82 is set as the open end 82a, and the open end 82a is set so that the liquid surface is opened to the atmosphere. The sample container 73 is configured to be immersed in the DNA solution of the sample container 73 containing the DNA solution.
The DNA solution is supplied into the flow path member 82 from the flow path 3 and the flow path 8
4, it is not necessary to suck the DNA solution, and a liquid supply mechanism (piping, syringe piston pump, solenoid valve, etc.) for introducing the DNA solution into the flow path 84 in the flow path member 82 is provided. This eliminates the need for the device, and makes it possible to reduce the size and cost of the device.

Furthermore, in the present embodiment, the step of cleaning the flow path 84 and the step of discharging the DNA solution can be performed in parallel in separate steps, so that a probe array can be formed in a short time. In addition, in the first embodiment, when the liquid discharge unit 71 moves, acceleration is applied to the washing water and the DNA solution held in the Teflon pipe 24 and the flow path 34, so that the meniscus in the nozzle 35 fluctuates. Although the ejection accuracy may be affected, according to this embodiment, the portion holding the DNA solution is always fixed, so that the meniscus does not fluctuate and the ejection accuracy can be improved. Will be possible. Further, in the present embodiment, since the liquid discharge head 79 is fixed and the membrane filter 74 is moved, the moving speed of the membrane filter 74 can be higher than in the first embodiment, and the microarray The creation time can be reduced as compared with the first embodiment.

In this embodiment, as shown in FIG. 9 (a), the flow path member 82 is set up substantially vertically on the liquid surface of the sample container 73. However, the present invention is not limited to this. For example, as shown in FIG. 9B, a configuration may be adopted in which the flow path member 82 is erected on the side surface of the sample container 73 so as to have a slight angle with respect to the horizontal plane. In that case, XYZ
The support direction of the liquid ejection head 79 by the robot 72 will also be changed to correspond.

[0045]

As described above, according to the present invention,
It is possible to provide a microarray manufacturing apparatus and a manufacturing method capable of manufacturing a microarray having a uniform spot diameter even when bubbles are mixed in the flow path.

[Brief description of the drawings]

FIG. 1 is a front view illustrating a configuration of a microarray manufacturing apparatus according to a first embodiment of the present invention.

FIGS. 2 (a) and 2 (b) are a top view and a detailed view of a portion B, respectively, showing the configuration of the microarray manufacturing apparatus according to the first embodiment of the present invention.

FIG. 3 is a perspective view of a liquid discharge unit mounted on the microarray manufacturing apparatus of the first embodiment.

FIGS. 4A to 4E are views for explaining a liquid discharge operation by a liquid discharge head of the liquid discharge unit of the first embodiment.

FIG. 5 is a diagram illustrating a drive waveform of a laminated piezoelectric element of the liquid ejection head according to the first embodiment.

FIG. 6 is a front view illustrating a configuration of a microarray manufacturing apparatus according to a second embodiment of the present invention.

FIG. 7 is a top view illustrating a configuration of a microarray manufacturing apparatus according to a second embodiment of the present invention.

FIG. 8 is a diagram illustrating a detailed configuration of a discharge unit according to a second embodiment.

FIG. 9A is a cross-sectional view of a liquid ejection head according to a second embodiment, and FIG. 9B is a diagram illustrating a modification thereof.

FIG. 10 is a perspective view for explaining a conventional method for manufacturing a microarray.

11 is a sectional view taken along line AA of FIG.

[Explanation of symbols]

 1, 2 Microarray manufacturing apparatus 20 Liquid discharge head 24 Teflon piping 25 Syringe piston pump 26 Liquid supply tank 27 Solenoid valve 28 Water supply pump 29 Multilayer piezoelectric element (drive means) 30 Frame 31 Flow path member 33 Liquid introduction port 34 Flow path 35 nozzle 36 Straight part 37 Tapered part 38 Air layer 71 Liquid discharge unit 72 XYZ robot (relative moving means) 73 Sample container 74 Membrane filter (substrate) 75 Movable stage 76 Cleaning tank 77 Filter base 78 Discharge unit 79 Liquid discharge head 80 Unit base 81 Discharge head base 82 Flow path member 82a Lower end 83 Connecting member 84 Flow path 85 Nozzle 86 Tapered part 87 Straight part 88 Base

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) G01N 33/543 525 G01N 35/02 F 35/02 37/00 102 35/10 C12N 15/00 F 37 / 00 102 G01N 35/06 A F-term (Reference) 2G058 AA09 CC09 EA03 EA11 EB00 ED02 ED07 ED16 GA02 4B024 AA19 BA80 CA01 HA12 HA19 4B029 AA27 BB15 BB16 BB17 BB20 CC03 CC10 CC11 4B063 QA01 Q52 QA18Q52

Claims (4)

[Claims] 1. A microarray manufacturing apparatus for manufacturing a microarray by discharging a small amount of a liquid containing a probe capable of specifically binding to a target substance onto a substrate, wherein the liquid is held in a flow path inside the microarray. A flow path member having a discharge port at one end of the flow path; a driving unit for moving the flow path member back and forth in a discharge direction; and a relative position of the substrate and the flow path member in a plane orthogonal to the discharge direction. Relative movement means for changing the positional relationship, wherein the liquid held in the flow path member is discharged from the discharge port by moving the flow path member forward and backward along the discharge direction by the driving means. Microarray manufacturing apparatus characterized by the following: 2. A liquid container for supplying the liquid to the flow path member, the liquid container storing the liquid such that the liquid surface is open to the atmosphere. The other end of the passage is an open end, the open end is immersed in the liquid in the liquid container, and the liquid is supplied from the liquid container into the flow path member by capillary action. The microarray manufacturing apparatus according to claim 1, wherein: 3. The microarray manufacturing according to claim 2, wherein the relative moving means is configured to change the relative positional relationship between the substrate and the flow path member by moving the substrate. apparatus. 4. A method for manufacturing a microarray, comprising manufacturing a microarray using the microarray manufacturing apparatus according to claim 1.